Introduction: Pyridoxine-dependent epilepsy is a rare cause of seizures in childhood. The diagnosis is made upon clinical criteria, that in many cases are never met. Therefore, epidemiologic data on pyridoxine dependency are scarce. Objectives: To study the epidemiology of pyridoxine dependent epilepsy in The Netherlands, and to determine whether the diagnosis is based upon the appropriate criteria. Methods: Nationwide all departments of paediatrics (n=113) and of paediatric or neonatal neurology (n=17) were asked to report cases of pyridoxine-dependent seizures. Birth incidences were calculated using national data on life births from 1991 to 2003. Results: Response was received from 67% of paediatric departments, including all university hospitals and 94% of child neurology departments. Thirteen patients were reported. Five definite (38%), two probable (15%), and four possible cases (31%) were identified. Two cases (15%) did not meet criteria for either of these groups. The birth incidence was 1:396 000 for definite and probable cases and 1:252 000 when possible cases are included. Conclusions: Thus far, epidemiologic data on pyridoxine dependent seizures were only available from the UK and Ireland. A higher incidence was found in The Netherlands, in accordance to earlier suggestions of a regional difference. The study shows that the diagnosis is often made without performance of a formal trial of withdrawal. We want to underline the importance of confirming the diagnosis, concerning the consequences as for individual prognosis, the potential side effects of prolonged pyridoxine substitution, and the possibility of treating the mother in case of future pregnancies.